The Phene PlateTM systemFingerprinting of bacteria in microplates

PhPlateMicroplate techniques

PhPlateMicroplate techniques

Principles of bacterial typingPhenotyping - analysis of the expression of the bacterial cell

Genotyping - analysis of the bacterial genome

Kemotyping - analysis of the contents of the bacterial cell or the cell envelope

PhPlateMicroplate techniques

Different kinds of bacterial fingerprintsDNA fingerprintBiochemical fingerprint

PhPlateMicroplate techniques

Microbial ecologyMeasures diversities of microflorasMeasures stability of the microfloraMeasures similarity between different microflorasEpidemiologyCan be used to verify whether isolates are of different clonal origin Can be used as a pre-screening method prior to DNA fingerprintingThe PhenePlateTM system can e.g. be used forAreas of application

PhPlateMicroplate techniques

How can you trace bacteria in surface water, drinking water, or food1)Inoculate or 16 24 colonies of indicator bacteria per sample in PhenePlates 2)Read the result and analyze with the PhP-software

PhPlateMicroplate techniques

Tracing sources of bacterial pollutions in surface waterHuman fecalmaterialIndustrialeffluentsAnimal waste

PhPlateMicroplate techniques

Tracing bacteria in consumers waterLow diversity =Regrowth within the systemHigh diversity =Inadequate purification

PhPlateMicroplate techniques

Tracing bacteria in food

PhPlateMicroplate techniques

How can you - simply detect a food borne outbreak caused by e.g. ETEC or EHEC ?

1)Inoculate 8 or 16 E. coli colonies per sample (including food) in RE PhenePlates 2)Read the result and analyze with the PhP-software

PhPlateMicroplate techniques

If no common strain no outbreak !3)Look at the dendrogram for a common (outbreak) strain. For confirmation, analyze only one ore a few isolates belonging to the common strain for appropriate properties Food borne outbreak

PhPlateMicroplate techniques

How can you- find out whether the same strain of Enterococcus sp. causes the UTI of Mrs. Svenson in ward A as in Mrs. Olsen in ward B ?PS: It works with Escherichia coli etc. as well1.Inoculate the strains in the FS-PhenePlate 2.Read and analyze the result with the PhP-software3.Look at the computer generated list of identical strains

Srain namePhP typeSvensonA1SvensonB1OlsenA1OlsenB1Control-12Control-23

PhPlateMicroplate techniques

How can you

- find out whether the Klebsiella strains in the neonatal ward originate from a common source ? 1)Inoculate the strains in the PhP-48 plates2)Read and analyze the result with the PhP-software3)Look at the dendrogram for a common outbreak strain

PhPlateMicroplate techniques

How can I

- reduce the number of isolates that need to be DNA typed? 1)Inoculate all isolates into the PhP plates 2)Read and analyze the result with the PhP-software3)Look at the dendrogram, and select only those isolates that could not be differentiated by PhP for DNA typing

PhPlateMicroplate techniques

Example: Clustered PhP data and PFGE patterns for VRE isolatesHow does PhP typing perform in comparison to DNA typing?

PhPlateMicroplate techniques

Checking bacterial cultures for purityPick one pure colony and one pooled colony (or streak) for PhP typing Different biochemical fingerprints low correlations - indicate unpure culturesIdentical biochemical fingerprints high correlations - indicate pure cultures D1 col r= 0.997 99 22 2 24 24 23 22 22 23 9 8 6 D1 pool 99 21 2 23 23 23 22 21 23 10 9 6 D3 col r= 0.907 99 22 8 8 9 13 21 21 24 23 10 14 D3 pool 99 21 5 8 8 13 21 22 21 14 7 14 D4 col r= 0.995 99 23 3 22 23 24 23 22 24 17 24 6 D4 pool 99 23 2 22 24 24 23 22 24 17 22 6 D6 col r= 0.706 99 23 2 23 6 23 23 22 24 12 8 6 D6 pool 99 24 5 9 8 8 23 22 24 11 10 8

PhPlateMicroplate techniques

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